Background Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is a tool that has revolutionised clinical microbiology and has recently been described as an innovative and effective approach to arthropod identification. Methods In this study, mosquitoes were captured in Vietnam using four different methods (human landing catch, CDC light traps, BG-Sentinel traps, animal-baited net traps). A total of 4215 mosquitoes were captured and morphologically identified as belonging to three genera: Aedes, Anopheles and Culex. We randomly selected 1253 mosquitoes, including 662 specimens of 14 Anopheles species, 200 specimens of two Aedes species and 391 morphologically unidentified Culex specimens, for molecular and MALDI-TOF MS analysis. The DNA from 98 mosquitoes (69 Anopheles specimens, 23 Culex specimens and six Aedes sp. specimens) was subjected to molecular analysis, either to confirm our morphological identification or the MALDI-TOF MS results, as well as to identify the Culex species that were morphologically identified at the genus level and to resolve the discrepancies between the morphological identification and the MALDI-TOF MS identification. Results High-quality MS spectra were obtained for 1058 of the 1253 specimens (84%), including 192/200 for Aedes, 589/662 for Anopheles and 277/391 for Culex. The blind test showed that 986/997 (99%) of the specimens were correctly identified by MALDI-TOF MS, with log score values ranging from 1.708 to 2.843. Eleven specimens of Culex could not be identified based on morphological features, MALDI-TOF MS or molecular analysis. Conclusions This study enabled us to identify several species of mosquitoes from Vietnam using MALDI-TOF MS, and to enrich our database of MALDI-TOF MS reference spectra. Graphical Abstract

Mosquito-borne diseases pose a significant threat to humans in almost every part of the world. Key factors such as global warming, climatic conditions, rapid urbanisation, frequent human relocation, and widespread deforestation significantly increase the number of mosquitoes and mosquito-borne diseases in Vietnam, and elsewhere around the world. In southeast Asia, and notably in Vietnam, national mosquito control programmes contribute to reducing the risk of mosquito-borne disease transmission, however, malaria and dengue remain a threat to public health. The aim of our review is to provide a complete checklist of all Vietnamese mosquitoes that have been recognised, as well as an overview of mosquito-borne diseases in Vietnam. A total of 281 mosquito species of 42 subgenera and 22 genera exist in Vietnam. Of those, Anopheles, Aedes, and Culex are found to be potential vectors for mosquito-borne diseases. Major mosquito-borne diseases in high-incidence areas of Vietnam include malaria, dengue, and Japanese encephalitis. This review may be useful to entomological researchers for future surveys of Vietnamese mosquitoes and to decision-makers responsible for vector control tactics.